本文選題：成骨細(xì)胞　切入點(diǎn)：膠原液晶　出處：《暨南大學(xué)》2015年碩士論文

【摘要】：天然骨組織是一種具有完美分級(jí)結(jié)構(gòu)的、天然的復(fù)合材料。在成骨細(xì)胞作用下,沿C軸取向生長(zhǎng)的納米羥基磷灰石(Hydroxyapatite,HA)沿著膠原纖維的平行方向有序聚集形成具有特殊的高級(jí)自組裝結(jié)構(gòu),從而使天然骨具有優(yōu)良的抗壓或抗張等物理性質(zhì)及生物性能。在體外成骨細(xì)胞的作用下,模擬天然骨成骨的生理環(huán)境,從而系統(tǒng)得研究骨組織形成的本質(zhì)及結(jié)構(gòu)與性能的關(guān)系,對(duì)設(shè)計(jì)高性能的仿骨材料具有重要意義。為了模擬天然骨組織的礦化過程,本文以甘油磷酸鈣(Glycerol phosphate calucium salt/Calcium Glycerophosphate,GPC)為鈣磷源,以I型膠原(Type I collagen,Col I)為大分子基質(zhì)材料,在堿性磷酸酶(Alkaline phosphatase,ALP)或前體成骨細(xì)胞的作用下,通過Sol-Gel相轉(zhuǎn)變礦化方法和細(xì)胞誘導(dǎo)礦化的方法,構(gòu)建了納米HA/Col復(fù)合材料。一方面從仿生角度出發(fā),制備出膠原液晶膜:采用偏光顯微鏡、掃描電鏡和原子力顯微鏡研究膠原膜的液晶織構(gòu)和拓?fù)浣Y(jié)構(gòu)。通過XRD、SEM、TEM觀察和EDS等測(cè)試對(duì)GPC-Col-ALP的礦化體系對(duì)綜合測(cè)評(píng),探究探討鈣磷無機(jī)礦物在液晶態(tài)膠原基質(zhì)上沉積的過程和機(jī)理。另一方面通過MTT法、電鏡觀察、細(xì)胞骨架熒光染色、骨鈣素染色、馮.卡門染色、堿性磷酸酶觀察研究GPC礦化體系對(duì)小鼠前體成骨細(xì)胞(MC-3T3-E1)細(xì)胞的增殖、生長(zhǎng)形貌及細(xì)胞骨架的影響。結(jié)果表明,在ALP能夠解離GPC上的磷酸酯鍵,從而使GPC解析出PO43-和Ca2+,在弱堿性和體溫的條件下生成羥基磷灰石,膠原具有促進(jìn)該體系礦化的作用。低濃度的甘油磷酸鈣礦化培養(yǎng)基具有促進(jìn)MC-3T3-E1細(xì)胞增殖生長(zhǎng)的功能,當(dāng)其濃度達(dá)到一定濃度15mmol/L后抑制細(xì)胞的增殖生長(zhǎng)。GPC能夠促進(jìn)成骨細(xì)胞分泌ALP,一定濃度范圍內(nèi),隨著GPC濃度的增大,ALP的活性增大,GPC在15mmol/L時(shí),ALP的活性最大,隨著GPC的濃度增大,ALP的活性逐漸減小。
[Abstract]:Natural bone tissue is a kind of natural composite material with perfect grading structure. Nano-hydroxyapatite HA, grown along C-axis orientation, aggregates in an ordered manner along the parallel direction of collagen fibers to form a special advanced self-assembly structure. So that the natural bone has excellent physical properties and biological properties, such as compressive or tensile resistance. Under the action of osteoblasts in vitro, the physiological environment of natural bone osteogenesis is simulated, and the nature of bone formation and the relationship between structure and performance are systematically studied. In order to simulate the mineralization process of natural bone tissue, the calcium Glycerol phosphate calucium salt/Calcium Glycerate Phosphate (GPC) was used as calcium and phosphorus source and type I collagen type I collagen I collagenesis (Col I) was used as macromolecular matrix material in order to simulate the mineralization process of natural bone tissue. Under the action of alkaline phosphatase Alkaline phosphatase (ALP) or precursor osteoblasts, Sol-Gel phase transformation mineralization and cell induced mineralization were used to construct HA/Col nanocomposites. The preparation of collagen liquid crystal membrane: the liquid crystal texture and topological structure of collagen membrane were studied by polarizing microscope, scanning electron microscope and atomic force microscope. The mineralized system of GPC-Col-ALP was comprehensively evaluated by TEM observation and EDS test. To explore the process and mechanism of calcium phosphate inorganic mineral deposition on liquid crystal collagen matrix. On the other hand, through MTT method, electron microscope observation, cytoskeleton fluorescence staining, osteocalcin staining, von Carmen staining, The effects of GPC mineralization system on the proliferation, morphology and cytoskeleton of mouse precursor osteoblast (MC-3T3-E1) cells were studied by alkaline phosphatase assay. The results showed that ALP could dissociate the phosphate bonds on GPC, so that GPC could dissociate PO43- and Ca2. Hydroxyapatite was formed under the condition of weak alkalinity and body temperature. Collagen promoted the mineralization of the system. The low concentration of calcium glycerophosphate mineralized medium could promote the proliferation and growth of MC-3T3-E1 cells. When the concentration of 15mmol/L reached a certain concentration, inhibiting the proliferation and growth of osteoblasts. GPC could promote the secretion of ALP by osteoblasts. In a certain concentration range, the activity of ALP increased with the increase of GPC concentration. The activity of GPC decreased with the increase of GPC concentration.
【學(xué)位授予單位】：暨南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R318.08

【共引文獻(xiàn)】

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